• Title/Summary/Keyword: In-vitro techniques

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New in vitro multiple cardiac ion channel screening system for preclinical Torsades de Pointes risk prediction under the Comprehensive in vitro Proarrhythmia Assay concepta

  • Jin Ryeol An;Seo-Yeong Mun;In Kyo Jung;Kwan Soo Kim;Chan Hyeok Kwon;Sun Ok Choi;Won Sun Park
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.3
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    • pp.267-275
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    • 2023
  • Cardiotoxicity, particularly drug-induced Torsades de Pointes (TdP), is a concern in drug safety assessment. The recent establishment of human induced pluripotent stem cell-derived cardiomyocytes (human iPSC-CMs) has become an attractive human-based platform for predicting cardiotoxicity. Moreover, electrophysiological assessment of multiple cardiac ion channel blocks is emerging as an important parameter to recapitulate proarrhythmic cardiotoxicity. Therefore, we aimed to establish a novel in vitro multiple cardiac ion channel screening-based method using human iPSC-CMs to predict the drug-induced arrhythmogenic risk. To explain the cellular mechanisms underlying the cardiotoxicity of three representative TdP high- (sotalol), intermediate- (chlorpromazine), and low-risk (mexiletine) drugs, and their effects on the cardiac action potential (AP) waveform and voltage-gated ion channels were explored using human iPSC-CMs. In a proof-of-principle experiment, we investigated the effects of cardioactive channel inhibitors on the electrophysiological profile of human iPSC-CMs before evaluating the cardiotoxicity of these drugs. In human iPSC-CMs, sotalol prolonged the AP duration and reduced the total amplitude (TA) via selective inhibition of IKr and INa currents, which are associated with an increased risk of ventricular tachycardia TdP. In contrast, chlorpromazine did not affect the TA; however, it slightly increased AP duration via balanced inhibition of IKr and ICa currents. Moreover, mexiletine did not affect the TA, yet slightly reduced the AP duration via dominant inhibition of ICa currents, which are associated with a decreased risk of ventricular tachycardia TdP. Based on these results, we suggest that human iPSC-CMs can be extended to other preclinical protocols and can supplement drug safety assessments.

Subtractive versus additive indirect manufacturing techniques of digitally designed partial dentures

  • Snosi, Ahmed Mamdouh;Lotfy, Shaimaa Mohamed;Thabet, Yasmine Galaleldin;Sabet, Marwa Ezzat;Rizk, Fardos Nabil
    • The Journal of Advanced Prosthodontics
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    • v.13 no.5
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    • pp.327-332
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    • 2021
  • PURPOSE. The purpose of this in vitro study was to evaluate the accuracy of digitally designed removable partial denture (RPD) frameworks, constructed by additive and subtractive methods castable resin patterns, using comparative 3D analysis. MATERIALS AND METHODS. A Kennedy class III mod. 1 educational maxillary model was used in this study. The cast was scanned after modification, and a removable partial denture framework was digitally designed. Twelve frameworks were constructed. Two groups were defined: Group A: six frameworks were milled with castable resin, then casted by the lost wax technique into Co-Cr frameworks; Group B: six frameworks were printed with castable resin, then casted by the lost wax technique into Co-Cr frameworks. Comparative 3D analysis was used to measure the accuracy of the fabricated frameworks using Geomagic Control X software. Student's t-test was used for comparing data. P value ≤ .05 was considered statistically significant. RESULTS. Regarding the accuracy of the occlusal rests, group A (milled) (0.1417 ± 0.0224) showed significantly higher accuracy than group B (printed) (0.02347 ± 0.0221). The same results were found regarding the 3D comparison of the overall accuracy, in which group A (0.1501 ± 0.0205) was significantly more accurate than group B (0.179 ± 0.0137). CONCLUSION. In indirect fabrication techniques, subtractive manufacturing yields more accurate RPDs than additive manufacturing.

Evaluation of intaglio surface trueness, wear, and fracture resistance of zirconia crown under simulated mastication: a comparative analysis between subtractive and additive manufacturing

  • Kim, Yong-Kyu;Han, Jung-Suk;Yoon, Hyung-In
    • The Journal of Advanced Prosthodontics
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    • v.14 no.2
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    • pp.122-132
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    • 2022
  • PURPOSE. This in-vitro analysis aimed to compare the intaglio trueness, the antagonist's wear volume loss, and fracture load of various single-unit zirconia prostheses fabricated by different manufacturing techniques. MATERIALS AND METHODS. Zirconia crowns were prepared into four different groups (n = 14 per group) according to the manufacturing techniques and generations of the materials. The intaglio surface trueness (root-mean-square estimates, RMS) of the crown was measured at the marginal, axial, occlusal, and inner surface areas. Half of the specimens were artificially aged in the chewing simulator with 120,000 cycles, and the antagonist's volume loss after aging was calculated. The fracture load for each crown group was measured before and after hydrothermal aging. The intaglio trueness was evaluated with Welch's ANOVA and the antagonist's volume loss was assessed by the Kruskal-Wallis tests. The effects of manufacturing and aging on the fracture resistance of the tested zirconia crowns were determined by two-way ANOVA. RESULTS. The trueness analysis of the crown intaglio surfaces showed surface deviation (RMS) within 50 ㎛, regardless of the manufacturing methods (P = .053). After simulated mastication, no significant differences in the volume loss of the antagonists were observed among the zirconia groups (P = .946). The manufacturing methods and simulated chewing had statistically significant effects on the fracture resistance (P < .001). CONCLUSION. The intaglio surface trueness, fracture resistance, and antagonist's wear volume of the additively manufactured 3Y-TZP crown were clinically acceptable, as compared with those of the 4Y- or 5Y-PSZ crowns produced by subtractive milling.

Microshear bond strength of dual-cure resin cement in zirconia after different cleaning techniques: an in vitro study

  • Atoche-Socola, Katherine Joselyn;Arriola-Guillen, Luis Ernesto;Lopez-Flores, Ana Isabel;Garcia, Isadora Martini;Huertas-Mogollon, Gustavo;Collares, Fabricio Mezzomo;Leitune, Vicente Castelo Branco
    • The Journal of Advanced Prosthodontics
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    • v.13 no.4
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    • pp.237-245
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    • 2021
  • PURPOSE. This study aimed to compare the microshear bond strength (µSBS) of dual-cure resin cement in CAD-CAM zirconia after different cleaning techniques. MATERIALS AND METHODS. Fifty discs of zirconia-based ceramic from Ivoclar Vivadent were embedded in acrylic resin. The discs were divided into five groups according to the cleaning methods used: Group 1: drying with spraying + sandblasting with Al2O3; Group 2: washed with water and dried with spraying + sandblasting with Al2O3;Group 3: washed with distilled water and dried with spraying + sandblasting with Al2O3 + zirconium oxide (Ivoclean); Group 4: washed with distilled water and dried with spraying + sandblasting with Al2O3 + potassium hydroxide (Zirclean); and Group 5: washed with distilled water and dried with spraying + sandblasting with Al2O3 + 1% NaClO. All of the groups were contaminated with artificial saliva for 1 minute and then cleaned. Statistical analyses were performed using ANOVA and Tukey's tests. RESULTS. There were statistically significant differences among all groups for µSBS (P < .05). The group treated with zirconium oxide (Group 3) showed the highest µSBS (18.75 ± 0.23 MPa). CONCLUSION. When applied to zirconia, the cleaning methods affected the bonding with resin cement differently.

Blood clot stabilization after different mechanical and chemical root treatments: a morphological evaluation using scanning electron microscopy

  • Stefanini, Martina;Ceraolo, Edoardo;Mazzitelli, Claudia;Maravic, Tatjana;Sangiorgi, Matteo;Zucchelli, Giovanni;Breschi, Lorenzo;Mazzoni, Annalisa
    • Journal of Periodontal and Implant Science
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    • v.52 no.1
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    • pp.54-64
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    • 2022
  • Purpose: This in vitro study was conducted to evaluate the effects of different debridement techniques and conditioning procedures on root surface morphology and blood clot stabilization. Methods: Two debridement techniques (curette [CU] vs. high-speed ultrasound [US]) and 2 conditioning procedures (ethylenediaminetetraacetic acid [EDTA] and phosphoric acid [PA]) were used for the study. Seven experimental groups were tested on root surfaces: 1) no treatment (C); 2) CU; 3) US; 4) CU+EDTA; 5) US+EDTA; 6) CU+PA; and 7) US+PA. Three specimens per group were observed under scanning electron microscopy (SEM) for surface characterization. Additional root slices received a blood drop, and clot formation was graded according to the blood element adhesion index by a single operator. Data were statistically analyzed, using a threshold of P<0.05 for statistical significance. Results: The C group displayed the most irregular surface among the tested groups with the complete absence of blood traces. The highest frequency of blood component adhesion was shown in the CU+EDTA group (P<0.05), while no differences were detected between the CU, US+EDTA, and CU+PA groups (P<0.05), which performed better than the US and US+PA groups (P<0.05). Conclusions: In this SEM analysis, EDTA and conventional manual scaling were the most efficient procedures for enhancing smear layer removal, collagen fiber exposure, and clot stabilization on the root surface. This technique is imperative in periodontal healing and regenerative procedures.

Studies on In Vivo Embryo Production by FSH-P and SUPER-OV in Korean Native Cattle H. The Factors Influencing In Vivo Embryo Production by Condition of Donors (한우에서FSH-P와 SUPER-OV에 의한 체내 수정란 생산에 관한 연구 II.공우란의 조건에 따른 체내 수정란 생산에 영향을 미치는 요인)

  • 김흥률;김덕임;박노형;김창근;정영채;윤종택;전광주
    • Journal of Embryo Transfer
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    • v.12 no.1
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    • pp.49-56
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    • 1997
  • This study was carried out to establish an effective and practical system for commercialization of embryo production techniques by analyzing several factors influencing in vivo embryo production on condition of donors treated FSH-P and SUPER-OV in Korean native cattle. In vivo embryos were flushed 226 times from 128 donors. The results obtained for the factors influencing in vivo embryo production by conditionof donors treated FSH-P and SUPER-OV were as follows :1. Age and parity of donors did not affect the transferable and freezable embryos among the treatments in FSH-P but the transferable and freezable embryos were decreased after 6 years old and 4th parity in SUPER-OV(P<0.01). 2. The production of embryos on the weight of donors was higher in over 400kg of body weight in FSH-P(P<0.01) and was higher in over 450kg than 400~450kg of body weight in SUPER-OV(P<0.05). For FSH-P embryo production was better responded in 350~450kg of body weight with 30~32mg doses, and showed a better result in over 450kg body weight with 32~34mg doses.(Key Words : in vivo embryo, donors, FSH-P SUPER-OV)

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Studies on the Factors Influencing In Vitro Embryo Production in Korean Native Cattle (한우의 체내 수정란 생산에 영향을 미치는 요인에 관한 연구)

  • 김홍률;김덕임;박노형;원유석;김창근;정영채;이규승;서길웅;박창식
    • Korean Journal of Animal Reproduction
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    • v.21 no.1
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    • pp.9-18
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    • 1997
  • These studies were carried out to establish an effective and practical system for commercialization of embryo production techniques by analyzing several factors influencing in vivo embryo production in Korean native cattle. In vivo embryos were flushed 226 times from 128 donors. The results obtained in studies on the factors influencing in vivo embryo production were as follows : 1. There were no significant differences in the number of total recovered, fertilized, transferable and freezable embryos among the hormone doses(FSH-P, 28∼34mg; SUPER-OV, 75IU) used for superovulation. However, over 30mg doses of FSH-P showed a slightly higher effect than others. 2. There were slight decrease in the number of fertilized, transferable and freezable embryos in 3 times repeated superovulation. But there were no significant differences among 1, 2 and 3 times repeated superovulations. 3. Age of donors did not affect the number of transferable and freezable embryos, but the number of fertilzed embryos were highest in 2∼3 years old donors and were lowest in 8∼9 years old donors(P<0.05). 4. Season had a significant effect on the production of embryos(P<0.05). the embryo production, and the number of fertilized, transferable and freezable embryos were most effective in summer and follow by spring, autumn and winter. 5. The number of transferable and freezable embryos was highest in donors flushed at 7∼8 days after estrus(P<0.05).

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Staphylococcus saprophyticus and Escherichia coli: Tracking from sperm fertility potential to assisted reproductive outcomes

  • Ghasemian, Fatemeh;Esmaeilnezhad, Shahin;Moghaddam, Mohammad Javad Mehdipour
    • Clinical and Experimental Reproductive Medicine
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    • v.48 no.2
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    • pp.142-149
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    • 2021
  • Objective: Bacteriospermia and urogenital infections are common problems in male infertility. This study aimed to evaluate the effects of bacteriospermia on sperm parameters and clinical outcomes in semen samples infected with two common bacteria (Staphylococcus saprophyticus and Escherichia coli) in northern Iran. Methods: Microbiological tests were performed to isolate and identify organisms from 435 semen samples from infertile couples. Semen samples were assessed according to the World Health Organization criteria. The protamine status, chromatin structure, chromatin condensation, and acrosome reaction of sperm and assisted reproductive outcomes were determined in couples with different male infertility factors. Results: Among the total cases, the two most prevalent pathogens were considered: S. saprophyticus (38.2%) and E. coli (52.9%). In the semen samples infected with E. coli, the spontaneous acrosome reaction and abnormal chromatin condensation were more common (p<0.05). Significant increases in abnormal chromatin condensation and deprotamination were seen in the presence of S. saprophyticus. In washed semen, tight adhesion between the sperm midpiece and S. saprophyticus was observed. There was also a significant decrease in the fertilization rate using semen samples infected with S. saprophyticus and E. coli during in vitro fertilization cycles (p<0.001). In addition, the presence of S. saprophyticus and E. coli in semen samples was associated with a lower likelihood of clinical pregnancy in couples with various factors of male infertility. Conclusion: Poor results of assisted reproductive techniques may be correlated with semen samples infected with two common bacteria in northern Iran.

Characterization of ginsenoside compound K loaded ionically cross-linked carboxymethyl chitosan-calcium nanoparticles and its cytotoxic potential against prostate cancer cells

  • Zhang, Jianmei;Zhou, Jinyi;Yuan, Qiaoyun;Zhan, Changyi;Shang, Zhi;Gu, Qian;Zhang, Ji;Fu, Guangbo;Hu, Weicheng
    • Journal of Ginseng Research
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    • v.45 no.2
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    • pp.228-235
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    • 2021
  • Backgroud: Ginsenoside compound K (GK) is a major metabolite of protopanaxadiol-type ginsenosides and has remarkable anticancer activities in vitro and in vivo. This work used an ionic cross-linking method to entrap GK within O-carboxymethyl chitosan (OCMC) nanoparticles (Nps) to form GK-loaded OCMC Nps (GK-OCMC Nps), which enhance the aqueous solubility and stability of GK. Methods: The GK-OCMC Nps were characterized using several physicochemical techniques, including x-ray diffraction, transmission electron microscopy, zeta potential analysis, and particle size analysis via dynamic light scattering. GK was released from GK-OCMC Nps and was conducted using the dialysis bag diffusion method. The effects of GK and GK-OCMC Nps on PC3 cell viability were measured by using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide assay. Fluorescent technology based on Cy5.5-labeled probes was used to explore the cellular uptake of GK-OCMC Nps. Results: The GK-OCMC NPs had a suitable particle size and zeta potential; they were spherical with good dispersion. In vitro drug release from GK-OCMC NPs was pH dependent. Moreover, the in vitro cytotoxicity study and cellular uptake assays indicated that the GK-OCMC Nps significantly enhanced the cytotoxicity and cellular uptake of GK toward the PC3 cells. GK-OCMC Nps also significantly promoted the activities of both caspase-3 and caspase-9. Conclusion: GK-OCMC Nps are potential nanocarriers for delivering hydrophobic drugs, thereby enhancing water solubility and permeability and improving the antiproliferative effects of GK.

Medical Application of Synchrotron Radiation in Japan

  • Toyofuku, Fukai;Tokumori, Kenji;Hyodo, Kazuyuki;Ando, Masami;Uyama, Chikao
    • Proceedings of the Korean Society of Medical Physics Conference
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    • 2002.09a
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    • pp.32-34
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    • 2002
  • Over the past two decades there has been a tremendous growth in the number of synchrotron radiation facilities in the world and also in Japan. The high flux and brightness radiation which derive from the third generation low emittance rings provide an ideal source for many applications in the medical sciences. The application of synchrotron radiation to medical imaging started in the early 80's in U.S.A, followed by European countries such as Germany and Russia. In Japan, researchs on intravenous coronary angiography started in 1884 at the Institute for High Energy Phisics(KEK) in Tsukuba. At present, it is the only application of syncrotron radiation which is at the stage of human study. In '90s, newer techniques such as phase and refraction contrast imaging appeared which are at the in vitro or animal study stage. Various types of x-ray CT have also been developed for three-dimensional imaging of the subjects. The present status of medical applications of synchrotron radiation in Japan is reviewed.

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